A thermostable 5-enolpyruvylshikimate-3-phosphate synthase from Thermotoga maritima enhances glyphosate tolerance in Escherichia coli and transgenic Arabidopsis
文献类型： 外文期刊
作者： Wang, Lijuan 1 ; Peng, Rihe 1 ; Tian, Yongsheng 1 ; Gao, Jianjie 1 ; Wang, Bo 1 ; Yao, Quanhong 1 ;
作者机构： 1.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai Key Lab Agr Genet & Breeding, 2901 Beidi Rd, Shanghai, Peoples R China
关键词： 5-Enolpyruvylshikimate-3-phosphate synthase; Thermotoga maritima; Hyperthermophile; Glyphosate tolerant; Transgenic Arabidopsis
期刊名称：EXTREMOPHILES （ 影响因子：2.395； 五年影响因子：2.668 ）
ISSN： 1431-0651
年卷期： 2019 年 23 卷 6 期
页码：
收录情况： SCI
摘要： 5-Enolpyruvylshikimate-3-phosphate synthase (EPSPS) overexpression, attempting to provide excess EPSPS to combine with glyphosate, is one way to improve glyphosate resistance of plants. The EPSPS in extremophiles which is selected by nature to withstand the evolutionary pressure may possess some potential-specific biological functions. In this study, we reported the cloning, expression and enzymatic characterization of a novel Class II EPSPS AroA(T. maritima) from Thermotoga maritima MSB8. The enzyme showed low sequence identities with other EPSPSs, and was one of the most thermostable EPSPSs so far, which showed the optimum enzyme activity at 80 degrees C. The enzyme maintains the activity below 50 degrees C and in a wide range of pH 4.0-10, which indicated its stability under rough environment, especially in tropical regions and alkaline soil. Excellent K-i/K-m value of AroA(T. maritima) suggested that the enzyme showed powerful competitive binding capacity of PEP over glyphosate and high glyphosate tolerance. Furthermore, aroA(T. maritima) gene was transformed into Arabidopsis thaliana. The transgenic lines were resistant to 15 mM glyphosate, which proved the application value in the cultivation of glyphosate-tolerant plants.
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